In vitro culture as a source of biodiversity for soybean breeding

The paper shows the effectiveness of methods of somaclonal variability in combination with multiple individual selection for creating new soybean varieties characterized by early ripeness and increased seed productivity for sub-arid regions of Russia and Kazakhstan with cold continental climate. Soybean breeding material was created using biotechnology methods combined with multiple individual selection. Higher regeneration potential of soybean tissues of SibNIIK 315 variety compared to Omskaya-4 variety was revealed. The ability to regenerate is associated with the adaptability of SibNIIK 315 variety, which is conﬁ rmed by the wider area of its cultivation in various ecological and geographical conditions in ﬁ ve regions of Russia and Kazakhstan. Tissues of apexes and cotyledonary nodes of R21 somaclonal line differed from the original SibNIIK 315 variety by the increased activity of callus formation, regeneration and growth of shoots. Tissues of regenerated plants passed repeatedly through the regeneration cycle according to the protocol of recurrent regeneration. Due to this, high morphogenic activity of R21 tissues can be the result of autoselection in vitro. The analysis of phenotypic variations in early generations of soybean somaclones showed an asymmetric distribution of deviations by the main breeding and marker traits: duration of vegetation, plant height and seed productivity, as well as dependence of distribution on weather conditions. In a dry year most somaclonal lines had a higher seed productivity, whereas in favourable conditions of a humid summer only every third line was superior to the original variety. This indicates increased levels of ontogenetic adaptation of somaclones in extreme drought conditions.

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Stimulation of Stem Callus and Regeneration of Ficus elastica "Decora" and from Apical Tips in vitro

Journal of Biotechnology Research Center ◽

10.24126/jobrc.2013.7.2.257 ◽

2013 ◽

Vol 7 (2) ◽

pp. 5-12

Author(s):

Ragad M. Abdullah ◽

Mazahim K. AL-Mallah

Keyword(s):

Callus Formation ◽

Potassium Nitrate ◽

Shoot Tips ◽

Peat Moss ◽

Ms Medium ◽

Ability To Regenerate ◽

Calli Cultures ◽

Clear Increase ◽

Stimulation Of

Calli cultures of stems and leaves explants excised from field -grown rubber, Ficus elastica Decora, plants were formed on agar-solidified Murashige and Skoog (MS) medium.The results proved that MS medium supplemented with 1.0 mg L-1 benzyl adenin (BA) and 0.8mg L-1 2,4-dichloro-phenoxy acetic acid (2,4-D) was suitable to stimulate stem’s callus at ratio 87.5% . Whereas supplementation of MS medium with 0.5 mg L-1 of both BA and Indole -3-butyric acid (IBA)encouraged callus formation to reach76.6%. Leaves showed responses for callus initiation up to 75% at the same media. Stem calli showed limited ability to regenerate shoots on both agar solidified MS medium containing 1.0 mg L-1 2,4-D and kinetin (kin) 0.5 mg L-1 and MS medium supplemented with 0.5 mg L-1 of both BA and IBA. Transferring of shoots to differentiation medium (MS+0.5 mg L-1 BA+ 0.1 mg L-1 IBA) stimulated the growth and elongation of these shoots. Shoots were transferred to agar-solidified MS medium free from growth regulators failed to form roots. Because of the less number of shoots, other rooting media were not tested. The data showed, that shoot tips succeeded to regenerate shoots when they were cultured in different MS. The results proved clear increase in chlorophyll and protein content of the rubber shoots as compared with content of field grown rubber plants. It was noticed that agar-solidified MS medium supplemented with 4.0 mg L-1 BA was considered the optimum medium for shoots regeneration. All plants regenerated from shoot tips were readily rooted in agar-solidified MS medium with increasing Potassium Nitrate KNO3 from 1900 mg L-1 to 2000 mg L-1, and at the same medium supplemented with 3.0 mg L-1 IBA and 1.0 mg L-1 BA. All these plants were successfully acclimated and transferred to peat moss.

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Stabilization of soft wheat selection material which is the result of crossing with wild relatives

Faktori eksperimental'noi evolucii organizmiv ◽

10.7124/feeo.v28.1380 ◽

2021 ◽

Vol 28 ◽

pp. 83-87

Author(s):

I. S. Zambriborshch ◽

O. L. Shestopal ◽

T. P. Nargan ◽

M. S. Chekalova

Keyword(s):

Winter Wheat ◽

Anther Culture ◽

Callus Formation ◽

Wild Relatives ◽

Soft Wheat ◽

Ability To Regenerate ◽

Androgenesis In Vitro ◽

Callus Regeneration ◽

Soft Winter Wheat

Aim. Testing the haploproduction ability of 30 hybrids of winter soft wheat. Methods. In vitro culture of isolated anthers of wheat. The percentage of callus and regeneration of green plants for each genotype calculated as a percentage of the planted anthers. Results. The differences in the frequency of callus induction and the ability to regenerate plants in the process of androgenesiss in vitro of winter soft wheat were detected. The microspores of 17 of 30 hybrids formed callus by in vitro anther culture were shown. The intensity of one process was different: more than half of the genotypes (18 pcs.) were characterized by a low percentage of callus (from 0.10 to 1.0%), 6 genotypes - medium (from 1.0 to 3.0%), and three - high (4.36%; 15.11% and 15.81%, respectively). Conclusions. Genotype-specific of microspores morphogenetic reactions of soft winter wheat in the process of androgenesis in vitro were revealed Samples P26 and P27 showed the highest level of callus formation. The 10 green regenerating plants were obtained. Keywords: hybrids, soft winter wheat, anther culture in vitro, callus, regeneration.

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Efficiency of in vitro culture techniques applied to soybean (Glycine max (L.) Merr.) accessions from the VIR collection

PROCEEDINGS ON APPLIED BOTANY GENETICS AND BREEDING ◽

10.30901/2227-8834-2021-4-137-142 ◽

2021 ◽

Vol 182 (4) ◽

pp. 137-142

Author(s):

E. S. Korshikova ◽

K. M. Ershova ◽

Yu. A. Moksheninova ◽

Yu. V. Ukhatova

Keyword(s):

Callus Formation ◽

Research Trend ◽

Culture Techniques ◽

New Genes ◽

Wide Range ◽

Ability To Regenerate ◽

Presidential Decree ◽

Genetic Technologies ◽

Adverse Environmental Conditions

Using a wide range of modern biotechnologies and genetic techniques to study plant germplasm accessions held by VIR makes it possible to procure valuable results, required for the development of new high-yielding cultivars adapted to adverse environmental conditions and possessing specified technological properties, particularly to identify and mark new genes and alleles useful for plant breeding. This research trend is in line with Presidential Decree No. 680 “Concerning the development of genetic technologies in the Russian Federation”. Soybean is among the key crops in agricultural production, but the use of next-generation breeding tools to obtain new soybean cultivars with desired properties is still limited. Successful application of novel methods also requires new approaches to studying soybean accessions, specifically their ability to regenerate and produce calluses for subsequent inclusion in biotechnological programs.Ten soybean accessions of various origin, contrasting in ripening schedules, were selected to study the possibility of effective introduction into in vitro culture and further assessment of their ability to produce calluses and regenerate in in vitro culture. The work included evaluating the effects of different seed sterilization techniques (one-step sterilization, using a commercial bleach, and two-step one, combining the impacts of a chlorine-containing preparation and hydrogen peroxide), types of explants (epicotyls, hypocotyls, cotyledon nodes, and cotyledon leaf segments), and phytohormone composition of nutrient medium: (1) MS + 1.13 mg/L BAP + 0.5 mg/L HA, and (2) MS +1 mg/L BAP + 0.1 mg/L IAA).The assessment results showed that the option of two-step seed sterilization was the most effective for soybean at the stage of in vitro culture initiation, while hypocotyls, epicotyls, and cotyledon nodes had the highest callus formation ability in both types of nutrient media.

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FACTORS AFFECTING THE REGENERATION OF PEPPER (CAPSICUM ANNUUM L.)

HortScience ◽

10.21273/hortsci.25.9.1120 ◽

1990 ◽

Vol 25 (9) ◽

pp. 1120G-1120

Author(s):

J. L. Jacobs ◽

C. T. Stephens

Keyword(s):

Growth Hormone ◽

Silver Nitrate ◽

Callus Formation ◽

Leaf Explants ◽

Regeneration System ◽

Factors Affecting ◽

Nitrate Concentrations ◽

Leaf Differentiation ◽

Whole Plants

Several growth hormone combinations and silver nitrate concentrations were examined for their effect on regeneration of different pepper genotypes. Primary leaf explants from in vitro seedlings were cultured on a revised Murashige and Skoog medium supplemented with auxin, cytokinin and 1.6% glucose. Combinations of different concentrations of indole-3-acetic acid (IAA), 0-5 mg/l, and 6-benzylaminopurine (BAP), 0-5 mg/l, were tested to determine the most effective medium for shoot primordium formation. Experiments with IAA and BAP did not result in a specific growth hormone combination appropriate for regeneration of all genotypes tested. Of the silver nitrate concentrations tested, 10 mg/l resulted in the best shoot and leaf differentiation and reduced callus formation. Differences in organogenic response of individual genotypes were evaluated on a single regeneration medium. Whole plants were regenerated from 11 of 63 genotypes examined. Based on these experiments, a reproducible regeneration system for pepper was developed with a total of 500 plants regenerated to date.

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Utilization of Different Seedling Explants for in Vitro Propagation of Hibiscus syriacus

HortScience ◽

10.21273/hortsci.33.3.478e ◽

1998 ◽

Vol 33 (3) ◽

pp. 478e-479

Author(s):

M.M. Jenderek ◽

A.J. Olney

Keyword(s):

Callus Formation ◽

Adventitious Bud ◽

Root Explants ◽

Petiole Explants ◽

Bud Formation ◽

Hypocotyl Explants ◽

Leaf Petiole ◽

Adventitious Bud Formation ◽

Hibiscus Syriacus

Hibiscus syriacus is a difficult species in micropropagation due to its endogenous contamination and recalcitrant shoot formation; therefore, studies on using explants other than shoot tip or axillary buds of growing shrubs were initiated. Three different seedling fragments (root, hypocotyl, and leaf petiole) from aseptically germinated seedlings of hibiscus (var. Aphrodite) were evaluated for adventitious bud formation, shoot and leaf development. The explants were cultured on McCown's woody plant basal salt medium supplemented with KNO3 (800 mg/L), adenine sulfate (80 mg/L) and MS vitamins containing BA or 2iP or TDZ at 0.5, 1.0, 2.2, 4.4 and 10 mM. Adventitious buds were present on all of the three different explants grown on medium containing TDZ; however, the most abundant bud formation, with many small leaves originating from callus was observed on hypocotyl explants cultured on medium with 1 mM of TDZ. Petiole explants were the most frequent to develop short shoots (≈15 mm) and one to nine leaves without callus formation, where 70% of hypocotyl and the root explants formed leaves originating from callus. Callus was induced on all explant types regardless of the level or type of cytokinin used. However, the number of shoots produced by any explant type was low, petioles cultured on 0.5 and 1mM of TDZ were the most suitable material for non-callus shoot development in H. syriacus. Hypocotyl explants proved to be an excellent source for adventitious bud formation but their ability to develop shoots needs to be investigated.

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Intensification de la régénération du pois (Pisum sativum L.), par le thidiazuron, via la formation de structures caulinaires organogènes

Canadian Journal of Botany ◽

10.1139/b97-053 ◽

1997 ◽

Vol 75 (3) ◽

pp. 492-500 ◽

Cited By ~ 6

Author(s):

Delphine Popiers ◽

Frédéric Flandre ◽

Brigitte S. Sangwan-Norreel

Keyword(s):

Pisum Sativum ◽

In Vitro Regeneration ◽

Naphthaleneacetic Acid ◽

Cotyledonary Nodes ◽

Pisum Sativum L ◽

Embryo Axes ◽

Initial Medium ◽

Second Wave ◽

Mature Seeds

In vitro regeneration of pea (Pisum sativum L.), a regeneration recalcitrant legume, was optimised using thidiazuron. Buds were initiated from the meristems of the cotyledonary nodes of embryo axes, isolated from mature seeds, and subcultured on Murashige and Skoog medium supplemented with 13.3 μM 6-benzylaminopurine, 16.1 μM α-naphthaleneacetic acid, and 0.2 μM 2,3,5-triiodobenzoic acid. Proliferation of buds was preceded by the formation of white nodular-like protrusions. These structures were cut transversally in fine slices and subcultured on the same medium or in presence of thidiazuron that produces a second wave of secondary budding. The best results (90–110 buds per expiant) were obtained with 10 μM thidiazuron. The capacity of regeneration was genotype independent and reproducible. Buds elongated on the initial medium, then formed roots in presence of 5.37 μM α-naphthaleneacetic acid. and developed into viable plants. Key words: Pisum sativum L., regeneration, meristems, embryo axes, thidiazuron.

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Somaclonal variation on in vitro callus culture potato cultivars

Horticultura Brasileira ◽

10.1590/s0102-05362004000200027 ◽

2004 ◽

Vol 22 (2) ◽

pp. 300-304 ◽

Cited By ~ 28

Author(s):

Patricia N. Bordallo ◽

Derly H. Silva ◽

José Maria ◽

Cosme D. Cruz ◽

Elizabeth P. Fontes

Keyword(s):

Somaclonal Variation ◽

Culture Media ◽

Callus Formation ◽

Potato Cultivars ◽

Synthetic Seed ◽

Arbitrary Sequence ◽

Stem Explants ◽

Factors Affecting ◽

High Level

Synthetic seeds can be an alternative for those species in which botanical seeds are not viable. One of the major problems of in vitro plant cultivation is the high level of somaclonal variation. The most common factors affecting somaclonal variation are genotype, explant source, in vitro period and cultivation conditions in which the culture is established. In this work, calli were induced using leaf and stem explants of the commercial potato cultivars Achat, Baraka, Baronesa, Bintje, and Contenda in MS culture media supplemented with 1.65 mM of picloram and 11.5 mM of 2,4-D. Seventy and 90 days after induction, DNA samples of 40 calli were compared concerning the effects of the two explant (leaf and stem) and two growth regulator sources on five potatoes cultivars. A total of 20 arbitrary sequence primers were evaluated. The RAPD pattern generated by these primers suggested a high percentage of polymorphic fragments among the five genotypes, indicating a high level of genetic variation among cultivars. Cultivar Baronesa showed the highest number of polymorphic fragments for all treatments. The cultivar Contenda showed the smallest somaclonal variation, for most of the treatments, except for the treatment which consisted of stem explants, picloram (1.65 mM) application, and a 70-day period of callus formation. 'Contenda' is, therefore, the most suitable cultivar for synthetic seed production.

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Organ culture with black cottonwood: morphogenetic response of female catkin primordia

Canadian Journal of Botany ◽

10.1139/b72-200 ◽

1972 ◽

Vol 50 (7) ◽

pp. 1627-1631 ◽

Cited By ~ 15

Author(s):

K. S. Bawa ◽

R. F. Stettler

Keyword(s):

Organ Culture ◽

Vegetative Growth ◽

Callus Formation ◽

Populus Trichocarpa ◽

General Tendency ◽

Black Cottonwood ◽

Axillary Shoots ◽

Female Sex ◽

Female Catkin

Female catkin primordia of black cottonwood (Populus trichocarpa T. & G. ex Hook.) were cultured for 70 days on a modified Murashige and Skoog's (1962) medium in vitro. Explants 2–3 mm long, and with bud scales removed, gave the best results, many of them developing floral structures characteristic of the female sex. There was a general tendency to callus formation with increasing age of the culture, occasionally followed by a reversal to vegetative growth. Catkin primordia raised on Wolter's medium without auxin or kinetin, but with 6-benzylaminopurine, and at 250 ft-c for a 16-h photoperiod, proliferated axillary shoots in loco of pistils.

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Selectivity of auxin for induction and growth of callus from excised embryo of spring and winter wheat

Canadian Journal of Botany ◽

10.1139/b84-189 ◽

1984 ◽

Vol 62 (7) ◽

pp. 1393-1397 ◽

Cited By ~ 6

Author(s):

M. D. Zhou ◽

T. T. Lee

Keyword(s):

Winter Wheat ◽

Chinese Spring ◽

Shoot Growth ◽

Callus Formation ◽

Triticum Aestivum L ◽

Dichlorophenoxyacetic Acid ◽

Naphthaleneacetic Acid ◽

2,4 Dichlorophenoxyacetic Acid ◽

Methoxybenzoic Acid

The callus-promoting activity of most commonly known as well as some rarely tested auxins was compared with that of 2,4-dichlorophenoxyacetic acid (2,4-D) for in vitro culture of the excised embryo of spring and winter wheat (Triticum aestivum L.), cv. Chinese Spring and cv. Fredrick. Different auxins in a concentration range from 1 to 50 μM showed widely different activities. Also the two wheat cultivars responded differently to the auxins. When rapid callus formation with limited root growth was used as the basis for comparison, 2-(2-methyl-4-chlorophenoxy)propionic acid (2-MCPP), α-naphthaleneacetic acid, 3,6-dichloro-2-methoxybenzoic acid (dicamba), 4-amino-3,5,6,trichloropicolinic acid (picloram), γ-(2,4-dichlorophenoxy)butyric acid, 2,4,5-trichlorophenoxyacetic acid, and 2,4,5-trichlorophenoxypropionic acid, in the order of effectiveness, were superior to 2,4,-D for callus induction from the embryo of 'Chinese Spring,' although the concentration required was higher than that of 2,4-D. For the winter wheat 'Fredrick,' however, only picloram, dicamba, and 2-MCPP performed as well as 2,4-D. All auxins tested promoted shoot growth; 2-methyl-4-chlorophenoxypropionic acid was most effective for 'Chinese Spring,' whereas picloram was most effective for 'Fredrick.'

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